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Search Results: 1 - 10 of 59 matches for " UTUT WIDYASTUTI "
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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.
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.
Physiological and Biochemical Responses to Aluminum Stress in the Root of a Biodiesel Plant Jatropha curcas L.
RADITE TISTAMA,UTUT WIDYASTUTI,DIDY SOPANDIE,AKIHO YOKOTA
HAYATI Journal of Biosciences , 2012,
Abstract: We investigated J. curcas responses to aluminum stress, histochemically and biochemically. Histochemical stainings were observed to analysis aluminum accumulation, lipid peroxidation and the loss of plasma membrane integrity on the surface and tissue of the root apex. Enzymatic analysis was conducted to measure malate content in leaf, root and malate efflux in the medium. We used M. malabathricum as a comparison for Al-tolerance plant. J. curcas root elongation was inhibited by 0.4 mM AlCl3, while M. malabathricum root elongation was inhibited by 0.8 mM AlCl3 treatment. Inhibition of root elongation has high correlation with Al accumulation in the root apex, which caused lipid degradation and cell death. Generally, malate content in J. curcas leaf and root was higher than that in M. malabathricum. In the contrary malate efflux from the root into the medium was lower. J. curcas root has a different pattern compared to M. malabathricum in malate synthesis and malate secretion when treated with a different Al concentration. We categorized J. curcas acc IP3 as more sensitive to aluminum than M. malabathricum.
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.
Diversity of SCAR Markers of Pyricularia grisea Isolated from Digitaria ciliaris Following Cross Infection to Rice
SRI LISTIYOWATI,UTUT WIDYASTUTI,GAYUH RAHAYU,ALEX HARTANA
Microbiology Indonesia , 2011, DOI: 10.5454/mi.5.1.1
Abstract: Cross infection of Pyricularia grisea from grass to rice and vice versa has been reported, but genetic changes are not known yet. This research aimed at estimating the possibility of the genotype alteration in P. grisea dc4 isolated from Digitaria ciliaris, following cross infection to either rice cv. Kencana bali, Cisokan, and IR64 or Panicum repens, Cynodon dactylon, Digitaria sp., and Ottochloa nodosa. The genotypes were analyzed by employing three SCAR markers, Cut1; PWL2; and Erg2. The results indicated that the dc4 was only able to infect Kencana bali, Cisokan, and P. repens. The dc4 had only two out of three SCAR markers, Cut1 and Erg2. Host shift was followed by genotype alteration in two loci of SCAR. Isolates derived from lesions on Kencana bali (dc4-kb) and Cisokan (dc4-c) of the dc4 infection, both lost their Cut1 and gained PWL2. On the contrary, there was no genotype alteration from dc4 to isolate derived from P. repens of dc4 infection (dc4-pr. Neither the isolate dc4-kb that was cross-inoculated to Cisokan nor the dc4-c that was cross-inoculated to Kencana bali showed SCAR marker change. In comparison, race 173 isolate and those derived from Kencana bali and Cisokan did not show genotype alteration. All had two out of three SCAR markers, PWL2 and Erg2. The isolate 173 was adapted to rice. This indicated that genotype diversity of the dc4 might arise following host shift from grass to rice.
ISOLATION, CLONING AND CHARACTERIZATION OF ACTIN-ENCODING cDNAs FROM Jatropha curcas L. IP-2P
Ratna Yuniati,Utut Widyastuti1,2,,Didy Sopandie,Akiho Yokota
Makara Seri Sains , 2011,
Abstract: Actin is a major component of the plant cytoskeleton, so all cells contain this protein. Actin is expressed constitutivelyand is involved in basic housekeeping functions required for cell maintenance. Because of this, it has been frequentlyused as an internal control to normalize changes in gene expressions analysis. Actually, the information of nucleotidesequence of actin gene of Jatropha curcas L. population IP-2P from Indonesia is not available yet. The objective of thisresearch was to isolate, clone and characterize cDNA of actin genes of J. curcas IP-2P. Three partial actin genesequences had been successfully isolated by PCR using total cDNA as template, and actin primer designed fromconserved region of Arabidopsis thaliana. Nucleotide sequence analysis showed that the length of JcACT fragment is610, 534, and 701 bp encoding 203, 177, and 234 amino acids respectively. Local alignment analysis based on mRNAsequences shows that JcACT fragment shares 98% similarity with actin mRNA of Hevea brasiliensis and 99% withactin mRNA of Ricinus communis. Based on deduced amino acid sequence, JcACT is 100% identical to actins fromPrunus 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.
Feeding Rate of Soil Animals in Different Ecosystems in Pati, Indonesia
RAHAYU WIDYASTUTI
HAYATI Journal of Biosciences , 2006,
Abstract: The feeding activity of soil animals was measured by using bait lamina test in three main ecosystems, i.e. the teak forest, home garden and rainfed paddy field. Two additional ecosystems in rainfed paddy field, i.e. the old (permanently established bund around paddy fields) and new bunds were examined as well. Three blocks of bait-lamina sticks (each block consisting of 16 individual sticks) were exposed at each location. The bait lamina were retrieved from the soil after two days and visually assessed. Each hole is designated as “fed” (perforated) or “non-fed” hole. The feeding rate is measured as the absolute number of “fed” holes. Soil animals in the old bunds showed the highest feeding activity (55.20%), followed by home garden (39.10%), rainfed paddy field (16.50%), teak forest (15.60%), and new bund (7.80%). The frequency of animals attack to the bait strips also indicated the similar pattern as their feeding activity, i.e. high in the old bunds (0.90), followed by home garden (0.70), teak forest (0.40), new bunds (0.40) and rainfed paddy field (0.30), respectively.
Physical Interactions between Yeast Pichia guilliermondii and Post-Harvest Fruit Pathogen Penicillium expansum
SRI WIDYASTUTI
HAYATI Journal of Biosciences , 2008,
Abstract: Attachment of yeast cells or bacteria on fungal hyphae have been observed in various antagonisms between microorganisms. Physical interactions between yeast Pichia guilliermondii and postharvest fruit pathogen Penicillium expansum in culture were studied in detail using light and transmission electron microscope to give better understanding on their mode of antagonism. Both organisms were co-cultured for 24-hr on potato dextrose agar. Light microscopy observations on the co-culture showed that the yeast cells attached firmly on the fungal hyphae. This attachment was inhibited by several substances such as enzymes degrading protein (protease or trypsin), a respiration inhibitor (sodium azide), an acid (hydrochloric acid) or an alkali (sodium hydroxide). Although autoclaved hyphae did not affect the attachment, but boiled enzymes and autoclaved yeast cells totally abolished the attachment. These evidences suggested that the attachment might be an active process mediated by certain protein from live yeast cells. Transmission electron micrographs on the ultrastructure of the co-culture revealed that the hyphae showed abnormalities in their structure and organelles, and a degree of obvious damage. Physical interactions observed in this study could be contributed to the mechanism of antagonism between P. guilliermondii and P. expansum.
Fermentasi Silase dan Manfaat Probiotik Silase bagi Ruminansia
Y. Widyastuti
Media Peternakan , 2008,
Abstract: Forage conservation has long been a part of the agricultural scene in some countries in the world. Ensilage is a preservation method for moist forages that is based on natural lactic acid fermentation under anaerobic conditions. There are six phases which occur during ensilage, storage and feed-out of the fermented forages. The technology of silage making is not popular in Indonesia, although ensilage may successfully occurs in tropical area including Indonesia. The reason may be due to limited information available regarding ensilage for the farmers. This review covered silage fermentation process and probiotics effect of feeding silage to the ruminants. The role of lactic acid bacteria is very important both from the preservation and antimicrobial points of view.
The Role of Lactic Acid Bacteria in Milk Fermentation  [PDF]
Yantyati Widyastuti, Rohmatussolihat  , Andi Febrisiantosa
Food and Nutrition Sciences (FNS) , 2014, DOI: 10.4236/fns.2014.54051
Abstract:

Species of lactic acid bacteria (LAB) represent as potential microorganisms and have been widely applied in food fermentation worldwide. Milk fermentation process has been relied on the activity of LAB, where transformation of milk to good quality of fermented milk products made possible. The presence of LAB in milk fermentation can be either as spontaneous or inoculated starter cultures. Both of them are promising cultures to be explored in fermented milk manufacture. LAB have a role in milk fermentation to produce acid which is important as preservative agents and generating flavour of the products. They also produce exopolysaccharides which are essential as texture formation. Considering the existing reports on several health-promoting properties as well as their generally recognized as safe (GRAS) status of LAB, they can be widely used in the developing of new fermented milk products.

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