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Isolation and Cloning of cDNA Fragment of Gene Encoding for Multidrug Resistance Associated Protein from M. affine.
Suharsono,Syarifin Firdaus,Utut Widyastuti Suharsono
Makara Seri Sains , 2008,
Abstract: Isolation and Cloning of cDNA Fragment of Gene Encoding for Multidrug Resistance Associated Protein from M. affine. M. affine can grow well in acid soil with high level of soluble aluminum. One of the important proteins in the detoxifying xenobiotic stress including acid and Al stresses is a multidrug resistance associated protein (MRP) encoded by mrp gene. The objective of this research is to isolate and clone the cDNA fragment of MaMrp encoding MRP from M. affine. By reverse transcription, total cDNA had been synthesized from the total RNA as template. The fragment of cDNA MaMrp had been successfully isolated by PCR by using total cDNA as template and mrp primer designed from A. thaliana, yeast, and human. This fragment was successfully inserted into pGEM-T Easy and the recombinant plasmid was successfully introduced into E. coli DH5α. Nucleotide sequence analysis showed that the lenght of MaMrp fragment is 633 bp encoding 208 amino acids. Local alignment analysis based on nucleotide of mRNA showed that MaMrp fragment is 69% identical to AtMrp1 and 63% to AtMrp from A. thaliana. Based on deduced amino acid sequence, MaMRP is 84% identical to part of AtMRP13, 77% to AtMRP12, and 73% to AtMRP1 from A. thaliana respectively. Alignment analysis with AtMRP1 showed that MaMRP fragment is located in TM1 and NBF1 domains and has a specific amino acid sequence QCKAQLQNMEEE.
EXPRESSION OF RESISTANCE OF SOYBEAN TO THE POD SUCKING BUG RIPTORTUS LINEARIS F. (HEMIPTERA: COREIDAE)
Suharsono,Liliek Sulistyowati
Agrivita : Journal of Agricultural Science , 2012,
Abstract: Factors involved in the mechanism of soybean resistance to pod sucking bug R. linearis were identified using resistant soybean genotypes, IAC-100, and IAC-80-596-2 and the susceptible variety, Wilis as a check. The role of trichomes in resistance was assayed removing trichomes from the pod shell, and seed coat and the resistance was determined based on the number of stylet punctures made by the bug. Seed of IAC-100 and IAC-596-2 that had longer, denser trichomes, higher crude fiber content and suffered fewer stylet punctures than Wilis. This suggested that denser and longer trichomes interfered with stylet piercing of the pod shell. When the trichomes of IAC-100 and IAC-596-2 were removed these genotypes were more susceptible to insect feeding. In further studies, replacement of IAC-100 and IAC-596-2 seed with seed of Wilis in the pods of resistant genotypes resulted less stylet punctures on the Wilis seed. It was concluded that denser and longer trichomes on pods along with harder pod shells acts as a physical barrier in antixenosis resistance of soybean to the pod sucking bug. Therefore, IAC-100, and IAC-596-2 genotypes have good potential for used as resistant parents in a soybean breeding program.
Isolation and Cloning of cDNA of Gene Encoding for Metallothionein Type 2 from Soybean [Glycine max (L.) (Merrill)] cv. Slamet
SUHARSONO,YASSIER ANWAR,UTUT WIDYASTUTI
Biodiversitas , 2009,
Abstract: Metallothionein has an important role in the detoxification of metal ions. It has a low molecular weight and contains cysteine-rich residue. The objective of this research is to isolate and clone the cDNA of gene encoding for metallothionein from soybean [Glycine max (L.) (Merrill)] cv Slamet (GmMt2). We had successfully isolated total RNA by reverse transcription and synthesized total cDNA from total RNA as template. cDNA of GmMt2 had been isolated from total cDNA by PCR. It was successfully inserted into pGEM-T Easy plasmid, and the recombinant plasmids were introduced into Escherichia coli strain DH5α. Sequence analysis by using T7 and SP6 primers showed that the length of PCR-isolated fragment is 257 bp containing 246 bp completed sequence of Mt2 cDNA encoding for 81 amino acids. Enzyme restriction analysis showed that GmMt2 does not contain any restriction sites found in the multi cloning sites of pGEM-T easy. Nucleotide and amino acid alignment analysis using BLAST program showed that GmMt2 is similar with completed cDNA of AtMt2A from Arabidopsis thaliana (L.) Heynh. Amino acid sequence analysis showed that the motifs of Cys sequence of GmMT2 are Cys-Cys, Cys-X-Cys, and Cys-X-X-Cys.
Morphological variation of Hoya multiflora Blume at different habitat type of Bodogol Research Station of Gunung Gede Pangrango National Park, Indonesia
SRI RAHAYU,MUHAMMAD JUSUF,SUHARSONO,CECEP KUSMANA
Biodiversitas , 2010,
Abstract: Rahayu S, Jusuf M, Suharsono, Kusmana C, Abdulhadi R. (2010) Morphological variation of Hoya multiflora Blume at different habitat type of Bodogol Research Station of Gunung Gede Pangrango Natonal Park, Indonesia. Biodiversitas 11: 187-193. Hoya multiflora Blume (Asclepiadaceae) is an Asiatic tropical ephypitic shrub which has been utilized as ornamental plant and reported to possess medicinal properties. The aim of this study was to evaluate the morphological variation of Hoya multiflora populations at the different habitat types of Bodogol Research Station of Gunung Gede Pangrango National Park in Indonesia. We collected 48 samples from three sub populations with six different habitat types. Morphological variation was found in stem, leave, and inflorescence. According to the discriminant and cluster analysis, the 48 samples were separated into three groups at 12 % dissimilarity. The groups were determined by canopy cover degree.
Isolation of cDNA Fragment of Gene Encoding for Actin from Melastoma malabthricum.
Saleha Hannum,Kinya Akashi,Utut Widyastuti Suharsono,Alex Hartana
Makara Seri Sains , 2010,
Abstract: Isolation of cDNA Fragment of Gene Encoding for Actin from Melastoma malabthricum. M. malabathricumgrows well in acidic soil with high Al solubility, thereby it can be used as a model plant for tolerance to aluminum andacid stresses. Actin is housekeeping gene used as an internal control for gene expression analysis. The objective of thisresearch was to isolate and clone the cDNA fragments of MmACT encoding for actin of M. malabathricum. Total RNAwas isolated and used as the template for cDNA synthesis by reverse transcription. Four cDNA fragments of MmACT,called MmACT1, MmACT2, MmACT3, and MmACT4, had been isolated and inserted into pGEM-T Easy plasmid.Nucleotide sequence analysis showed that the size of MmACT1 and MmACT2 is 617 bp, whereas MmACT3 andMmACT4 is 735 bp. The similarity among these four MmACT is about 78%-99% based on nucleotide sequence andabout 98%-100% based on amino acid sequence. Phylogenetic analysis based on amino acid sequence showed that at1% dissimilarity, the MmACT1, MmACT2, MmACT3 and the ACT5 Populus trichocarpha are clustered in one group,while the MmACT4 is grouped with ACT9 P. trichocarpa and ACT1 Gossypium hirsutum, and these two groups areseparated from actin group of monocotyledonous plants. The sequence of MmACT fragments were registered inGenBank/EMBL/DDBJ database with accession numbers AB500686, AB500687, AB500688, and AB500689.
ISOLATION AND CLONING OF cDNA OF GENE ENCODING FOR METALLOTHIONEIN TYPE 2 FROM MELASTOMA AFFINE
SUHARSONO,NIKEN TRISNANINGRUM,LULUT DWI SULISTYANINGSIH,UTUT WIDYASTUT
BIOTROPIA : the Southeast Asian Journal of Tropical Biology , 2009,
Abstract: Metallothionein is an important protein for detoxifying heavy metal ions. h is research was conducted to isolate and clone cDNA of gene encoding for metallothionein type 2 from Melastoma affi ne . Total RNA was isolated from young leaves. Total cDNA was synthesized from the total RNA by reverse transcription. h e MaMt2 cDNA was successfully isolated by PCR technique. h e MaMt2 cDNA was inserted into pGEM-T Easy and the recombinant plasmid was successfully introduced into Escherichia coli DH5 α . DNA sequencing analysis showed that this cDNA is full length consisting of 246 pb encoding 81 amino acid residues. h is cDNA is identical to mRNA of AtMt2 from Arabidopsis thaliana. It does not contain any restriction sites found in the cloning sites of pGEM-T Easy. h e deduced protein of MaMT2 contains 14 cysteine residues distributed in the Cys-Cys, Cys-X-Cys, and Cys-X-X-Cys motifs
In Situ Oxygen Dynamics in Coral-Algal Interactions
Daniel Wangpraseurt, Miriam Weber, Hans R?y, Lubos Polerecky, Dirk de Beer, Suharsono, Maggy M. Nugues
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0031192
Abstract: Background Coral reefs degrade globally at an alarming rate, with benthic algae often replacing corals. However, the extent to which benthic algae contribute to coral mortality, and the potential mechanisms involved, remain disputed. Recent laboratory studies suggested that algae kill corals by inducing hypoxia on the coral surface, through stimulated microbial respiration. Methods/Findings We examined the main premise of this hypothesis by measuring in situ oxygen microenvironments at the contact interface between the massive coral Porites spp. and turf algae, and between Porites spp. and crustose coralline algae (CCA). Oxygen levels at the interface were similar to healthy coral tissue and ranged between 300–400 μM during the day. At night, the interface was hypoxic (~70 μM) in coral-turf interactions and close to anoxic (~2 μM) in coral-CCA interactions, but these values were not significantly different from healthy tissue. The diffusive boundary layer (DBL) was about three times thicker at the interface than above healthy tissue, due to a depression in the local topography. A numerical model, developed to analyze the oxygen profiles above the irregular interface, revealed strongly reduced net photosynthesis and dark respiration rates at the coral-algal interface compared to unaffected tissue during the day and at night, respectively. Conclusions/Significance Our results showed that hypoxia was not a consistent feature in the microenvironment of the coral-algal interface under in situ conditions. Therefore, hypoxia alone is unlikely to be the cause of coral mortality. Due to the modified topography, the interaction zone is distinguished by a thicker diffusive boundary layer, which limits the local metabolic activity and likely promotes accumulation of potentially harmful metabolic products (e.g., allelochemicals and protons). Our study highlights the importance of mass transfer phenomena and the need for direct in situ measurements of microenvironmental conditions in studies on coral stress.
ASSESSING ECOLOGICAL RESILIENCE OF INDONESIAN CORAL REEFS
Imam Bachtiar1,2, Ario Damar1, Suharsono3, Neviaty P. Zamani4
Journal of Coastal Development , 2011,
Abstract: Ecological resilience is an important property of natural ecosystem to be understood in coral reef management. Resilience of Indonesian coral reefs was assessed using 2009 COREMAP data. The assessment used 698 data of line intercept transects collected from 15 districts and 4 marine physiographies. Resilience index used in the assessment was developed by the authors but will be published elsewhere. The results showed that coral reefs at western region had higher average resilience indices than eastern region, and Sunda Shelf reefs had higher resilience indices than coral reefs at Indian Ocean, Sulawesi-Flores, or Sahul Shelf. Four districts were found to have coral reefs with highest resilience indices, i.e. Bintan and Natuna (western region), and Wakatobi and Buton (eastern region). Raja Ampat had coral reefs with lower average resilience indices than that of Wakatobi. Uses of resilience index in coral reef management should be coupled with other information such as maximum depth of coral communities.
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