Kopi Luwak, an exotic Indonesian coffee, is made from coffee berries that have been eaten by the Asian palm civet ( Paradoxurus hermaphroditus ). Despite being known as the world's most expensive coffee, there is no reliable, standardized method for determining its authenticity. GC-MS-based multimarker profiling was employed to explore significant metabolites as discriminant markers for authentication. Extracts of 21 coffee beans ( Coffea arabica and Coffea canephora ) from three cultivation areas were analyzed and subjected to multivariate analyses, principal component analysis, and orthogonal projection to latent structures discriminant analysis. Citric acid, malic acid, and the inositol/pyroglutamic acid ratio were selected for further verification by evaluating their differentiating abilities against various commercial coffee products. The markers demonstrated potential application in the differentiation of original, fake Kopi Luwak, regular coffee, and coffee blend samples with 50 wt % Kopi Luwak content. This is the first report to address the selection and successful validation of discriminant markers for the authentication of Kopi Luwak.

Buasbuas (Premna pubescens) is a plant that is traditionally known to have medicinal properties. This study aims to determine the content of bioactive compounds contained in the petiole by Gas Chromatography Mass Spectrophotometer (GCMS) method. Preparation of ethanol extract of petiole using maceration method with 96% ethanol solvent. The study used gas chromatographic tools and mass spectra which were evaluated using MASSLAB program. The data obtained from the GCMS machine is then analyzed using the NCBI database pubchem software (https://pubchem.ncbi.nlm.nih.gov/). The results of this study indicate that the content of bioactive compounds on the petioles of buasbuas more than 50 libraries contains about 150 species of compounds with a range of RT and% area respectively 4.684 to 28.155 and 0.16 to 15.56%. The content of bioactive compounds shown this data indicates that very large potential of buasbuas plants to be explored and exploitation as a nutritious plant. The results of this study can be used as the foundation in the development program of the potential utilization of bioassemblance of buasbuas plants. In further research it is necessary to analyze the other parts of the plant and make comparisons to complete the available databases.

Polyhydroxybutyrate is a component of bioplas cs that is synthesized under the control of enzymes encoded by pha mul genes. The genes are naturally present in Ralstonia eutropha. However, the produc on of bioplas cs in bacteria is inefficient because the bacterial biomass is rela vely small compared with plants or fungi. As such, engineering techniques have been developed that enable pha genes to be inserted into plant biomass, and then be expressed in the biomass of the plant to produce polyhydroxybutyrate. The objec ves of this study were to transform the ssue of Jatropha curcas using the phaC gene (a pha gene), to regenerate the transformed plant, and to confirm the presence of the inserted genes with PCR. The gene c transforma on of J. curcas was mediated by Agrobacterium tumefaciens strain GV3101 containing pARTC by dipping the cotyledon ssue of J. curcas in a suspension of the bacterium for 30 min, followed by cocul va on for 3 d on Murashige and Skoog (MS) medium. The ssue was then placed on a selec on medium, i.e. MS medium containing 13.3 µM BAP and 0.05 µM IBA with the addi on of 20 mg/L kanamycin. The results showed that 12.35% of the ssue survived and regenerated into a shoot a er 1-2 months. Molecular analysis of the transformed ssue was performed using phaC and nptII primers, in order to detect the presence of the phaC and nptII genes. Specific bands were detected at 659 bp and 700 bp, corresponding to the nptII primer and phaC primer, respec vely.

The Effects of Aluminum Stress on Organic Acid Content of Lycopersicon esculentum Mill. cv. Intan Callus and Plantlet. Wening Enggarini and Erly Marwani. The purpose of this research was to evaluate the effects of Al stress on citric, malic and oxalic acid content of L. esculentum cv. Intan callus and plantlet, also aluminum content of L. esculentum plantlet. Callus was induced from cotyledone of L. esculentum on Murashige & Skoog (MS) media containing 10 -7 M NAA and 10 -6 kinetin. The callus was then transferred step wisely at 3 weeks interval to media containing 220, 275, 330, 385, 440, 550, 825, and 1100 μM AlCl 3 . The callus cultures on the control media and media with the addition of 550 μM AlCl 3 were able to regenerate and produce shoots after 8 passages of subculture. The shoots from media with the addition of 550 μM AlCl 3 were transferred into the media with addition of 825 μM AlCl 3 , then to the media with 1100 μM AlCl 3 . The High Pressure Liquid Chromatography (HPLC) analysis showed that Al stress callus and plantlets contained malic acid, but no citric and oxalic acid. The content of malic acid in callus decreased with increasing AlCl 3 concentration from 0 to 385 μM. On the other hand, the content of malic acid in callus increased with increasing AlCl 3 concentration from 440 μM to 1100 μM. Similarly, the content of malic acid in root increased with increasing concentration of AlCl 3 from 550 μM to 1100 μM. The result of Neutron Activation Analysis showed that Al content in root decreased as the amount of AlCl 3 increased in the media. These results suggested that L. esculentum callus and plantlet respond to the Al stress by producing higher amount of malic acid.

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Development the hairy root culture of A. paniculata was conducted for growing the hairy roots and production of andrographollide. Different strains of Agrobacterium rhizogenes (R-1000, A4, ATCC 15834), different types of explants (cotyledons, hypocotyls, and leaves) and different infection time of A. rhizogenes (1, 2, 3 days) were tested to induce hairy roots of A. paniculata. The results indicated that the best strain, type of explants and infection time for hairy roots induction were found in strain ATCC 15834, the explants of cotyledon and the 2 days of infection, respectively. The best medium for growing the hairy roots was liquid half strength MS medium with the addition of 5.0 µM IBA. The highest amount of andrographollide was observed in the medium with the addition of 5.0 µM IBA on the week of two, as much as 0.54%. Integration of T-DNA of A. rhizogenes in hairy roots was confirmed by polymerase chain reaction (PCR) analysis with specific primer for rolA and rolC genes of the plasmid. Visualization of the PCR products on agarose gel electrophoresis showed two fragments with lengths of 248 bp and 490 bp which corresponds to rolA and rolC genes from Ri plasmids of ATCC 15834.  Index Terms-A. paniculata; hairy roots; andrographollide;A. rhizogenes; rolA and rolC genes.

Objectives: To develop an efficient Botryococcus sp. for high biomass and oil productivity with a fatty acid profile suitable for biodiesel. Methods and analysis: The culture of Botryococcus sp.was maintained in the modified Chu-13 medium at 16:8 light and dark cycle with 5,200 lux light intensity at 25°C and aerated with 1.5 L of air per minute. The nitrogen concentrations of the media varied, 5, 28, 55 (control), 350 mg L −1 , with a constant concentration of phosphorous (44 mg L −1 ). For growth determination, Botryococcus sp. was harvested, dried, and weighed. Nitrogen consumption was measured by spectrophotometric methods. Oil was extracted from dried biomass of the algae using a soxhlet extractor (70°C, 4 hours), followed by methylation and analysis of fatty acids using gas chromatography. Findings: The results under these sets of experiments indicated that 350 mg N L −1 gave the maximal growth rate i.e. 0.24 g day −1 and the maximal biomass productivity i.e. 77.5 mg L −1 day −1 . On the other hand, the maximal oil productivity of 5.03 ± 0.10 mg L −1 d −1 was obtained from medium containing nitrogen 55 mgL −1 . The fatty acid profile of Botryococcus sp. oil showed a high amount of palmitic acid and oleic acid. Nitrogen concentrations of 5, 28, 55 (control) and 350 mg L −1 in the culture media resulted in 38.07%, 34.59%, 29.48%, 19.43% palmitic acid and 22.29%, 24.52%, 26.99%, 8.81% oleic acids respectively. These fatty acid profiles indicated that the Botryococcus sp. oil could be processed into biodiesel. Novelty: Growth, oil biomass content and fatty acid profiles of Botryococcus sp. cultures were affected by nitrogen concentration of the media. In this study, 55 mg N L −1 was the most efficient nitrogen concentration for maximal oil productivity with fatty acid content that met the requirements for conversion into biodiesel.

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