Indonesian Journal of Biotechnology
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Inter- and intraspecific variation of chloroplast mini- and microsatellites DNA in the four closed related Acacia species
No full textMini- and microsatellites of four Acacia species, A. aulacocarpa, A. auriculiformis, A. crassicarpa and A.mangium were investigated on four non-coding regions of cpDNA, the intron of trnL, and the intergenicspacers of trnL - trnP, trnD - trnY, and trnP – trnW. Nine single base substitutions and six informative miniandmicrosatellites were detected in the the four cpDNA non-coding regions. Based on the substitutionsand mini- and microsatellites, ten cpDNA haplotypes (A - J) could be distinguished. Acacia auriculiformispossessed fi ve haplotypes, A. aulacocarpa, four haplotypes, and A. crassicarpa, three haplotypes. All samplesof A. mangium possessed the same haplotype. Mini- and microsatellites recognized in this study can beused for species identifi cation of the four Acacia species. The ten haplotypes could divided the four speciesinto 2 groups, A. aulacocarpa-A.crassicarpa group and A. auriculiformis-A. mangium group. By developing thePCR-based markers based on the sequence information, many experiments can be carried out for the Acaciaimprovement programs
Molecular Identification of Phenol-Degrading and Biofi lm-Forming Bacteria from Wastewater and Peat Soil
No full textPhenol is hazardous aromatic pollutant which needs to be treated to reduce its hazardous effects.Bioremediation using bacteria which can form biofi lm offer an alternative wastewater treatment that is cheaperand environmentally safe. Eighteen strains of phenol-degrading and biofi lm-forming bacteria were isolatedfrom peat soil, also hospital and textile wastewater. Screening for phenol degradation ability of isolates wereperformed using Folin-ciocalteau reagent, while for biofi lm formation ability were performed using microtiterplate and crystal violet dye. Based on the ability to degrade phenol and to form biofi lm, four isolates (HP3,DOK135, DL120, andATA6) were choosen as phenol-degrading bacteria as well as biofi lm-forming bacteria.Based on phenotypic and genotypic characterization, isolate HP3 was highly similar to Rhodococcus equi strainDSM20307T, while DOK135 was highly similar to Enterobacter mori strain R18-2.The results also suggested thatDL120 and ATA6 could be classifi ed to the genus of Micrococcus and Bacillus respectivel
The Use of DNA Microsatellite Markers for Genetic Diversity Identification of Soybean (Glycine max (L) Meriil.) as a Supplementary Method in Reference Collections Management
No full textLarge number of new soybean varieties are mostly derived from crosses of elite genotypes resulted ina narrowing of both the genetic diversity and the phylogenetic relationship between soybean varieties. Thus,discrimination among soybean varieties is becoming more diffi cult, especially when morphological traits wereapplied. In Plant Variety Protection (PVP) system, new varieties of soybeans including granted PVP right, localand breeding varieties registered in PVP offi ce were frequently increased, implicate on increasingly the numberof soybean varieties collections. To assist the management of varieties collections, a standard fi ngerprinting datais further needed. In comparison to the management of plant collection in the fi eld, molecular marker systemswhich are rapid, reliable, informative and relatively simple are continually sought for practical applications ingermplasm conservation, management and enhancement. This study aimed to identify the genetic diversity andphylogenetic relationship of soybean varieties that have earned PVP Right as well as local varieties and breedingvarieties registered in the PVP offi ce using microsatellite or simple sequence repeats (SSR) markers.This study was conducted in Molecular Biology laboratory, Indonesian Center for Agricultural Biotechnologyand Genetic Resources Research and Development (ICABIOGRAD) Bogor, from February to May 2013. The datawere analyzed using the genetic analysis package NTSYSpc 2.02i and PowerMarker V3.25. The result showed arelatively narrow genetic diversity among 45 varieties of soybean analyzed in present study which were indicatedby the small number of genotypes and total number of alleles (NA), and the low value of gene diversity and PICvalues (<0.75). Cluster analysis showed that the grouping varieties are not related to morphological characters butrelated to phylogeny relationship between varieties. Despite the group of varieties were not clustered in accordancewith morphological characteristics, SSR marker can be a powerful tool for discriminating varieties, so that it couldbe useful for initial varieties identity in conjunction with genetic diversity analysis
Somatic embryogenesis of Sandalwood (Santalum album L.)
No full textSandalwood (Santalum album L.) is native species of Indonesia, especially in East Nusa Tenggara, is oneof the twenty two species of the genus Santalum in the world. Sandalwood is an important tree because it hashigh economic value can produce sandal oil these can be used for perfumes, cosmetics, pharmaceuticals, andare often used in religious ceremonies. In vitro particularly somatic embryogenesis has been widely appliedin the propagation of sandalwood. The Objective of this research is to obtain regeneration of sandalwoodthrough somatic embryogenesis using leaves explant from various clones. Medium for embryo induction is MS(Murashige and Skoog, 1962) solid medium containing treatment of 2,4-D (2,4-Dichlorophenoxyacetic acid)at various concentrations. To the media 0,15 mg /l kinetin, 40 g/l sucrose, and 2,5 g/l gelrite were added.Culture were incubated in the dark. Medium for Embryo development (maturation) is MS solid mediumcontaining treatment of BAP (Benzyl-amino-purine) at various concentrations. To the media 0,01 mg /l NAA(Napthalene-acetic-acid), 40 g/l sucrose, and 2,5 g/l gelrite were added. Culture were incubated in the light. Tostudy the specifi c structure of sandalwood somatic embryo early detection was conducted using histologicalanalysis. Results of anova showed that the clones, media, and interaction between clones with media did notsignifi cantly affect the development of sandalwood callus percentage. Results of anova showed that the clonesand BAP concentration signifi cantly effect to the embryo development of sandalwood
Polymorphism of Transcription Factor 7-Like 2 Gene and HOMA-β Level of Individuals With and Without Type 2 Diabetes Mellitus Family History
No full textFamily history has considered as a risk factor of type 2 diabetes. Transcription factor-7 like 2 (TCF7L2) hasrole to regulates insulin secretion and blood glucose homeostasis. The aim of current study was to determine thers7903146 polymorphism of TCF7L2 gene and homeostatic model assessment-β (HOMA-β) level on individual withand without type 2 Diabetes Mellitus (DM) family history. This work is a case-control study. Thirty six subjectswith type 2 DM family history and 36 subjects without type 2 DM family history were recruited. HOMA-βmeasure to analyze the insulin secretion. Polymorphisms of TCF7L2 gene was analyzed by using PCR-RFLPmethod. Statistical analysis was performed by using T-test, Mann-Whitney and Chi-square with signifi cancelevel 0.05. The frequency of the T allele of the cases were 4.2% and the controls were 2.8% (p=0.500). The oddratio was 0.649 (CI;95%:0.106-4.055). The HOMA-β levels of the cases were signifi cant low (132.56±62.48)compared with the controls (266.09±1.68) with p=0.000. The subjects with type 2 DM family history have asimilar frequency of having T alleles and CT/TT genotypes. The subjects with type 2 DM family history hassignifi cantly lower HOMA-β levels than subject without DM family history
The Phylogenetic Relationship Among Varieties of Lansium domesticum Correa Based on ITS rDNA Sequences
No full textLansium domesticum Corr. with vernacular name in Indonesian duku has been reported containingtherapeutic bioactive compounds, and some of these compounds shown to be potent antitumor, anticancer,antimalaria, antimelanogenesis, antibacteria, and antimutagenic activities. This plant is commonly known asduku, kokosan and langsat by the local community in Indonesia. The morphological appearance of all varieties isnearly the same, and identifi cation of the varieties is very diffi cult for growers. Variation of DNA sequences ofthe ITS (Internal transcribed spacer) region can be used as a molecular character to determine the phylogeneticrelationship of different varieties of L. domesticum. The aims of this study were to determine taxonomy status ofduku, kokosan, and langsat, also phylogenetic relationship among varieties of L. domesticum based on ITS rDNAsequencing. DNA was isolated from leaves of plant and then amplifi ed using F1 and R1 primers. Nucleotidesequences were identifi ed using Sequence Scanner Software Programm version 1.0, nucleotide sequences from18S, ITS1, 5.8S, ITS2 and 26S region, that has been mergered using EditSeq and SegMan in software Suite forSequence Analysis DNASTAR Lasergene DM version 3.0.25. The results of study showed that DNA fragmentsranging in size from 782-810 bp. Different pattern of DNA fragments indicated polymorphism among duku,kokosan, and langsat. Based on the results of the ITS rDNA sequencing and phylogenetic tree analysis. Itwas determined that Lansium and Aglaia are a separated genus with the similarity index value of 0.98. Duku,kokosan and langsat were divided into two cluster, namely cluster kokosan-langsat and cluster duku with thesimilarity index value of 0.996. Keywords : Phylogenetic relationship, ITS region, L. domesticum, duku, kokosan, langsa
Ethanol Fermentation on Mixed Sugars Using Mixed Culture of Two Yeast Strains
No full textThe objective of this study was to evaluate the use of mixed cultures of the recommended yeast strainsfrom a previous study on ethanol fermentation using a substrate mixture consisting of sucrose, glucose, andfructose. There were three mixed (combination) cultures namely OUT7096/OUT7913, OUT7096/OUT7921,and OUT7913/OUT7921. The fermentation medium contained sugar mixture consisting of glucose, fructose,and sucrose with a composition generally close to the composition of sugars in sweet sorghum juice. Overall,fermentation is carried out in two stages. First fermentation was performed using the three mixed culturesto determine the best combination based on the concentration of ethanol produced and the concentration ofresidual sugar. Second fermentation was conducted using the best mixed culture obtained from the fi rst stage.This second stage was intended to describe the pattern of the changes in the concentration of ethanol, sugarsand biomass during the fermentation progresses and to determine some kinetic parameters namely ethanolyield (Yp/s), growth yield (Yx/s) and specifi c growth rate (μ). The results of the fi rst fermentation showed thatthe best mixed culture was OUT7913/OUT7921 and the subsequent fermentation using this culture providethe highest ethanol yield (Yp/s) = 0.47 g.g-1 that was reached at 53rd hour, growth yield (Yx/s) = 0.425 g.g-1, andμ = 0.12 hour-1.Keywords : fermentation, ethanol, mixed culture, mixed suga
Improvement of Seed Orchard Management Based on Mating System of Cajuputi Trees
No full textBreeding plan of cajuputi in Indonesia is aimed to increase plantation productivity of oil yield and 1.8 cineole content. Seed orchard of cajuputi at Paliyan, Gunungkidul, established using selected and genetically improved materials, has been producing seeds for operational plantation. This seed orchard would perform optimally if the mating systems of all individuals contribute to the inheritance of all genetic potential of the offsprings. Therefore, investigation of the mating systems of cajuputi was indispensible. The study has been carried out on 10 selected mother trees and the 24 offsprings of each mother trees using 8 microsatellite markers of nuclear DNA, namely Hin-2 (100-132 bp), Hin-4 (79-114 bp), Hin-5 (128-148 bp), Hin-7 (136-224 bp), Sal-1 (93-99 bp), Sal-3 (118-219 bp), Xho-1 (96-111 bp) and Xho-4 (150-216 bp), respectively. The result showed relatively high genetic variation of the offspring (HE=0.602, HO=0.594) originated from parent trees in the seed orchard. Parent trees tend to outcross(tm=0.951, ts=0.806), although seeds originated from biparental inbred (tm – ts = 0.145) and correlated paternity(rp=0.098) have also been observed. This genetically viable population could maintain its reproduction fi tness forshort term and adapt to the dynamic environmental changes for long term. Key words: mating system, cajuputi, seed orchard, microsatellit
Phylogeny and Origin of Glucose-6-Phosphate Dehydrogenase (G6PD) Deficiency Mutations in Indonesia
No full textThe aim of this study is to analyze the relationship between the types of G6PD mutations found in Indonesia and the relationships of mutations found in Indonesia to those found in other countries. We summarize the distribution of G6PDs in West Indonesia and East Indonesia. Moreover, we use bioinformatics methods to construct phylogenetic trees and compare the sequences containing the regions amplifi ed by the commonly used PCR primer pairs. Previous work has shown that Mediterranean G6PD and Chinese CoimbraG6PDare distributed in West Indonesia, whilst G6PD mutations in East Indonesia are Jammu/ViangchanG6PD and Chinese Gaohe G6PD. G6PD Jammu/Viangchan was mostly distributed in Flores Island, East Indonesia along with G6PDGaohe. We constructed phylogenetic trees using the G6PD sequences from various regions in Indonesia and other countries. It appears from phylogenetic trees and percentages of identity that FloresIndonesian G6PD defi ciency (Jammu/Viangchan G6PD, originating in India) is 92.5% identical to the G6PD defi ciency of Chinese origin (GaoheG6PD). It was interesting to note that the genetic region containing the Javanese Indonesian G6PD defi ciency (MediterraneanG6PD, fi rst found in Italy) located in the western parts of Indonesia is closely related (99% identity) to the Chinese G6PD defi ciency(Coimbra G6PD). We concludethat G6PD mutations in West Indonesia are closely related to G6PD mutations from China. G6PD mutations in East Indonesia are also closely related to G6PD mutations from India and China, but more distantly, and to different types to those in West Indonesia. A prediction of protein structure was carried out which allowed visualization of the locations of mutation on the three dimensional structure of G6PD. Key words: G6PD, phylogeny, origin, genetic mutation
Comparative Analysis of Genes Induced by Respiratory Syncytial Virus and DsRNA in Human Epithelial Cells
No full textEpithelial cells are the primary target of respiratory viral infections and play a pivotal role in virusinducedlung infl ammation and in anti viral immune response. A common signal for the presence of viralinfections and induction of infl ammation is recognition of double stranded RNA (dsRNA). Thus far, therehas not been a high-throughput transcrptome analysis of RSV- or dsRNA-induced genes in primary humanbronchial epithelial cells (PHBE), nor there has been a comparison between dsRNA- and RSV-inducedgenes. To establish the transcriptome profi les and to determine the contribution of dsRNA in the inductionof infl ammation during respiratory virus infection, we compared the gene expression profi les of PHBE cellsthat were infected with Respiratory Syncytial Virus (RSV) or were treated with dsRNA. Our transcriptomeanalysis showed that RSV infection and and dsRNA treatment induced up-regulation of 2024 and 159 genesin PHBE respectively. Comparison of genes revealed that RSV and dsRNA commonly induced 75 genes inPHBE cells. The common up-regulated genes were functionally grouped in multiple response pathwaysinvolved in infl ammation and immune responses. Interestingly, there were several previously unreportedgenes that were up-regulated in primary human epithelial cells that are relevant to a TH2 allergic phenotype.This comparison of a high-throughput gene expression study offers a comprehensive view of transcriptionalchanges induced by dsRNA and RSV, and importantly compares dsRNA-induced genes with RSV-inducedgenes in PHBE cells. Keywords: RSV, dsRNA, transcriptome, immune response, infl ammatio